Graded fibronectin receptor aggregation in migrating cells

We have examined the distribution of the integrin fibronectin receptor in migrating NIH 3T3 fibroblasts to test the hypothesis that cell locomotion involves regional differences of adhesive receptor aggregation. A distinct asymmetry of fibronectin receptor aggregation was observed on the surface of migrating NIH 3T3 fibroblasts. Direct current electric fields were used to stimulate directional cell migration and thus allow the quantitative correlation of receptor asymmetries to the direction of cell locomotion. Digital particle analysis of fluorescent confocal micrographs demonstrates that the leading half of cells has a higher proportion of receptors contained in small clusters than does the trailing half. Conversely, larger receptor aggregates are more prevalent in the rear of the cell than in the front. We have also observed a gradient of extracellular fibronectin fibril assembly, similar to that described for the fibronectin receptor. Extracellular fibronectin appears in progressively larger fibrils across the ventral cell surface from front to rear, with dense meshworks often deposited as a trail behind the cell. Large fibronectin receptor clusters toward the rear of the cell generally do not correlate with focal contacts, and are thus most likely aggregated by cell surface-bound fibronectin fibrils and not by adhesion to the substratum. These results suggest that spatial variations in the degree of adhesive receptor aggregation are created in fibroblasts during the processes of migration and matrix synthesis.